初步探研栀子提取物对大鼠酒精性肝病血脂和TNF-α、IL-6变化的影响

发布时间：2018-05-31 06:15

本文选题：栀子提取物 + 酒精性肝损害　；参考：《遵义医学院》2014年硕士论文

【摘要】：目的：构建SD大鼠酒精性肝病的动物模型，并灌胃栀子提取物进行干预，然后通过测试实验大鼠的血清脂质总胆固醇(TC)、甘油三酯(TG)、谷丙氨酸氨基转移酶(ALT)、天冬氨酸氨基转移酶(AST)的水平，测定肝脏指数及肝脏组织中谷胱甘肽过氧化物酶(GSH-Px)、丙二醛(MDA)的含量，再观测肝脏组织肿瘤坏死因子-α (TNF-α)、白细胞介素-6(IL-6)蛋白表达和基因转录水平以及大鼠肝细胞脂肪变情况，以便探研酒精性肝病的病理机制及栀子提取物对其病变的影响。方法：五十只健康的SD雄性大鼠，随机分为五组（n=10），正常对照组（对照组）、酒精性肝病模型组（模型组）、栀子提取物低剂量组（低剂量组）、栀子提取物中剂量组（中剂量组）和栀子提取物高剂量组（高剂量组）。除了对照组大鼠给予普通饲料和清洁饮用水外，其余各组均每天给予高脂饲料和56%乙醇15ml/kg每天灌胃，各药物剂量组同时每天再添加低、中、高剂量的栀子提取物灌胃，，共连续饲养8周。然后分别麻醉动物，取材制备实验动物标本。（1）采取肝脏用电子天平称重，计算肝指数；（2）采用生化法检测血清的TC、TG、ALT和AST，并用试剂盒检测大鼠肝脏中GSH-Px和MDA的含量；（3）通过HE染色、免疫组化染色法分别用光镜观察肝组织的形态变化和TNF-α、IL-6的表达变化阳性指标；（4）提取肝脏组织的RNA，通过荧光定量RT-PCR法分别检测肝组织中TNF-α和IL-6mRNA的基因转录水平。结果：（1）动物用药8周后，计算大鼠肝指数，发现模型组较对照组的肝指数增高（P＜0.05），各药物组肝指数较模型组则明显降低（P＜0.05）。（2）模型组大鼠血清的TC、TG含量明显高于对照组（P＜0.05），各药物组与模型组比较，血清脂质明显降低（P＜0.05），模型组肝功能ALT、AST值明显较高于对照组（P＜0.05），各药物组肝功能指标则低于模型组（P＜0.05）；肝脏MDA含量模型组明显高于对照组（P＜0.05），各药物组MDA含量较模型组显著降低（P＜0.05）；模型组GSH-Px含量明显低于对照组（P＜0.05），药物组GSH-Px含量明显比模型组提高（P＜0.05）。（3）HE染色光镜下观察，各药物组的肝细胞脂肪变程度相比模型组大鼠而言有明显改善；免疫组化染色结果显示模型组较对照组大鼠肝脏组织的TNF-α、IL-6表达增多（P＜0.05），各药物组明显比模型组降低（P＜0.05）。（4）RT-PCR结果表明，模型组肝组织内TNF-α、IL-6mRNA的基因表达量均明显高于对照组，也高于各药物组（P＜0.05）。结论：（1）成功建立了SD大鼠酒精性肝脏损害的模型，并应用了栀子提取物来干预改善模型组大鼠的肝损害。（2）模型组大鼠的血清TC、TG、ALT、AST水平和MDA含量升高，GSH-Px含量下降；利用栀子提取物来进行药物干预，可以使血清TC、TG、ALT、AST水平和MDA含量降低，GSH-Px含量升高。（3）模型组大鼠肝组织的TNF-α、IL-6基因转录和蛋白表达升高，采用栀子提取物的药物干预可使其降低。总的来看，模型组SD大鼠的血清脂质的TC和TG水平升高,肝脏的ALT、AST和MDA水平升高，而GSH-Px含量降低，以及肝组织的TNF-α和IL-6基因转录表达增高，与酒精性肝脏损害有关；栀子提取物可能是通过下调TNF-α和IL-6转录表达、促进脂肪代谢、清除肝组织内自由基等，从而发挥保肝的治疗作用。
[Abstract]:Objective: to construct an animal model of alcoholic liver disease in SD rats, and to interfere with the extract of Gardenia jasminoides gardenia, and then test the level of serum lipid total cholesterol (TC), triglyceride (TG), alanine aminotransferase (ALT) and aspartate aminotransferase (AST), and determine the liver index and glutathione peroxy in the liver tissue. The content of chemical enzyme (GSH-Px) and malondialdehyde (MDA), the expression of tumor necrosis factor - alpha (TNF- alpha), the expression of interleukin -6 (IL-6) protein and gene transcription level, and the fatty change of rat liver cells, in order to explore the pathological mechanism of alcoholic liver disease and the effect of the extract of Gardenia jasminoides on its pathological changes.
Methods: Fifty healthy SD male rats were randomly divided into five groups (n=10), normal control group (control group), alcoholic liver disease model group (model group), low dose group of Gardenia extract (low dose group), middle dose group of Gardenia extract (medium dose group) and high dose group of Gardenia extract (high dose group). Except for control group, rats were given ordinary diet. And the other groups were given high fat diet and 56% ethanol 15ml/kg every day. Each drug dose group added low, medium and high doses of Gardenia extract for 8 weeks. Then the animals were reared for 8 weeks. Then, the animals were anaesthetized and prepared to prepare the experimental animal specimens. (1) the liver scales were weighed and the hepatic finger was calculated. (2) TC, TG, ALT and AST were detected by biochemical method, and the content of GSH-Px and MDA in rat liver was detected by the kit. (3) the morphological changes of liver tissue and the positive index of TNF- A and IL-6 expression were observed by HE staining and immunohistochemical staining method respectively. (4) the RNA of the liver tissue was extracted by the fluorescent quantitative RT-PCR method. The transcriptional levels of TNF- and IL-6mRNA genes were detected in liver tissues.
Results: (1) after 8 weeks of drug use, the liver index of rats was calculated and the liver index of the model group was higher than that of the control group (P < 0.05). The liver index of each group was significantly lower than that in the model group (P < 0.05). (2) the serum TC, TG content of the model group was significantly higher than that of the control group (P < 0.05). The serum lipids were significantly lower than those in the model group (P < < < < 0. 0.05) in the model group, the liver function ALT, AST value was significantly higher than the control group (P < 0.05), the liver function index of each drug group was lower than that of the model group (P < 0.05), the liver MDA content model group was significantly higher than the control group (P < 0.05), the MDA content in each drug group was significantly lower than the model group (P < 0.05), and the GSH-Px content in the model group was significantly lower than that of the control group (P < 0.05). The drug was significantly lower than the control group (P < 0.05). The content of GSH-Px in the group was significantly higher than that in the model group (P < 0.05). (3) under the HE staining light microscope, the fatty change degree of the liver cells in each drug group was significantly improved compared with the model group. The immunohistochemical staining results showed that the expression of TNF- a and IL-6 in the liver tissue of the model group increased (P < 0.05), and the drug groups were obviously compared with the model group. The group decreased (P < 0.05). (4) the results of RT-PCR showed that the gene expression of TNF- alpha and IL-6mRNA in the liver tissue of the model group was significantly higher than that in the control group, and was higher than that of the drug groups (P < 0.05).
Conclusion: (1) the model of alcoholic liver damage in SD rats was successfully established and the extract of Gardenia jasminoides was used to intervene to improve the liver damage in the model rats. (2) the serum levels of TC, TG, ALT, AST and MDA in the model group were increased and the content of GSH-Px decreased, and the serum TC, TG, ALT and AST levels could be made by using the extract of Gardenia jasminoides. And the content of MDA decreased and the content of GSH-Px increased. (3) the transcriptional and protein expression of TNF- alpha and IL-6 gene in the rat liver tissue of the model group increased, and the drug intervention by the extract of Gardenia could reduce it.
In total, the level of serum lipid TC and TG in the model group SD rats increased, the level of ALT, AST and MDA in the liver increased, but the content of GSH-Px decreased, and the transcriptional expression of TNF- A and IL-6 genes in the liver tissues increased, which was related to the alcoholic liver damage. The extract of Gardenia may be through down regulation of TNF- A and IL-6 transcription, promoting fat metabolism, and In addition to the free radicals in liver tissue, the therapeutic effect of liver protection is realized.
【学位授予单位】：遵义医学院
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R285.5

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